Requirement of Gαi1 and Gαi3 in interleukin-4-induced signaling, macrophage M2 polarization and allergic asthma response

Theranostics. 2021 Mar 4;11(10):4894-4909. doi: 10.7150/thno.56383. eCollection 2021.

Abstract

IL-4 induces Akt activation in macrophages, required for full M2 (alternative) polarization. We examined the roles of Gαi1 and Gαi3 in M2 polarization using multiple genetic methods. Methods and Results: In MEFs and primary murine BMDMs, Gαi1/3 shRNA, knockout or dominant negative mutations attenuated IL-4-induced IL4Rα endocytosis, Gab1 recruitment as well as Akt activation, leaving STAT6 signaling unaffected. Following IL-4 stimulation, Gαi1/3 proteins associated with the intracellular domain of IL-4Rα and the APPL1 adaptor, to mediate IL-4Rα endosomal traffic and Gab1-Akt activation in BMDMs. In contrast, gene silencing of Gαi1/3 with shRNA or knockout resulted in BMDMs that were refractory to IL-4-induced M2 polarization. Conversely, Gαi1/3-overexpressed BMDMs displayed preferred M2 response with IL-4 stimulation. In primary human macrophages IL-4-induced Akt activation and Th2 genes expression were inhibited with Gαi1/3 silencing, but augmented with Gαi1/3 overexpression. In Gαi1/3 double knockout (DKO) mice, M2 polarization, by injection of IL-4 complex or chitin, was potently inhibited. Moreover, in a murine model of asthma, ovalbumin-induced airway inflammation and hyperresponsiveness were largely impaired in Gαi1/3 DKO mice. Conclusion: These findings highlight novel and essential roles for Gαi1/3 in regulating IL-4-induced signaling, macrophage M2 polarization and allergic asthma response.

Keywords: Gαi1/3; IL-4; M2 polarization; allergic asthma response; signaling.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Asthma / immunology*
  • GTP-Binding Protein alpha Subunits, Gi-Go / genetics*
  • GTP-Binding Protein alpha Subunits, Gi-Go / immunology
  • Interleukin-4 / immunology*
  • Macrophages / immunology*
  • Mice
  • Mice, Knockout
  • Ovalbumin
  • Proto-Oncogene Proteins c-akt / metabolism
  • Respiratory Hypersensitivity / genetics*
  • Respiratory Hypersensitivity / immunology
  • TOR Serine-Threonine Kinases / metabolism

Substances

  • Gnai1 protein, mouse
  • Il4 protein, mouse
  • Interleukin-4
  • Ovalbumin
  • Proto-Oncogene Proteins c-akt
  • TOR Serine-Threonine Kinases
  • GTP-Binding Protein alpha Subunits, Gi-Go
  • Gnai3 protein, mouse